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HUDSON ALVES PINTO INFECÇÃO NATURAL DE Melanoides

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HUDSON ALVES PINTO INFECÇÃO NATURAL DE Melanoides 1 HUDSON ALVES PINTO INFECÇÃO NATURAL DE Melanoides tuberculata (MOLLUSCA: THIARIDAE) POR Centrocestus formosanus (TREMATODA: HETEROPHYIDAE) E POR Philophthalmus gralli (TREMATODA: PHILOPHTHALMIDAE) NO BRASIL Universidade Federal de Minas Gerais Instituto de Ciências Biológicas Belo Horizonte 2009 1 HUDSON ALVES PINTO INFECÇÃO NATURAL DE Melanoides tuberculata (MOLLUSCA: THIARIDAE) POR Centrocestus formosanus (TREMATODA: HETEROPHYIDAE) E POR Philophthalmus gralli (TREMATODA: PHILOPHTHALMIDAE) NO BRASIL Dissertação apresentada ao Programa de Pós-Graduação em Parasitologia, Instituto de Ciências Biológicas, Universidade Federal de Minas Gerais, como requisito parcial à obtenção do título de Mestre em Parasitologia. Área de Concentração: Helmintologia Orientador: Prof. Dr. Alan Lane de Melo Belo Horizonte 2009 2 043 Pinto, Hudson Alves. Infecção natural de Melanoides tuberculata (Mollusca: Thiaridae) por Centrocestus formosanus (Trematoda : Heterophyidae) e por Philophthalmus gralli (Trematoda : Philophthalmidae) no Brasil. [manuscrito] / Hudson Alves Pinto. – 2009. 99 f. : il. ; 29,5 cm. Orientador: Alan Lane de Melo. Dissertação (mestrado) – Universidade Federal de Minas Gerais, Departamento de Parasitologia. 1. Melanoides tuberculata. 2. Thiaridae - Teses. 3. Trematódeo - Teses. 4. Centrocestus formosanus. 5. Philophthalmus gralli. 6. Infecção - Teses. 7. Pampulha, Lagoa da (MG)- Teses. I. Melo, Alan Lane de. II. Universidade Federal de Minas Gerais. Departamento de Parasitologia. III. Título. CDU:576.88/.89 3 Trabalho desenvolvido no Laboratório de Taxonomia e Biologia de Invertebrados, Departamento de Parasitologia, Instituto de Ciências Biológicas, Universidade Federal de Minas Gerais. 4 Aos meus pais 5 AGRADECIMENTOS Ao professor Alan Lane de Melo, amigo e orientador, pela formação científica, disponibilidade e incentivo para a realização deste trabalho. Aos meus pais, Ozanan Pinto e Maria Lúcia Alves Pinto pela criação e apoio irrestrito à minha formação acadêmica e humana. À Priscila Amanda Alves Demétrio, futura esposa, pelo amor, carinho, convivência e conselhos que sem dúvida foram fonte de inspiração para as atividades científicas. Aos meus irmãos, Daniel Alves Pinto e Gabriela Aparecida Camargos Pinto pela amizade e companheirismo. A grande amiga Sirléia Aparecida Martins dos Reis e meu afilhado Christian Leandro dos Reis pelo carinho e apoio. A todos os familiares que de alguma forma incentivaram e contribuíram para minha formação. Ao Sr. Airton Lobo pela amizade, pelas conversas agradáveis e principalmente pelo excelente auxílio técnico e ensinamentos, de forma paciente, de todas as etapas necessárias à preparação do material de estudo. Ao grande amigo Vítor Luís Tenório Mati, pelo entusiasmo e interessantes discussões científicas compartilhadas durante a Pós-Graduação. Ao amigo Sandro Eugênio Gazzinelli pela disposição atenciosa para discutir assuntos diversos. Ao estimado amigo Gustavo José Caldas Vianna. Ao amigo Fernando Sérgio Barbosa pelo auxílio em algumas coletas malacológicas e em atividades laboratoriais. Ao estudante Hígor Rodrigues Duarte pela companhia e auxílio prestado durante algumas coletas malacológicas. 6 A Dra. Wanderlany Amâncio Martins pela consideração e amizade. Ao Dr. Marco Antônio Andrade de Souza pela amizade e incentivo aos estudos na área de malacologia. Aos amigos do Laboratório de Esquistossomose, Zenir de Souza, Selma Fernandes de Souza, Florence Mara Rosa, José Carlos Reis e em especial ao Sr. Alberto Geraldo dos Santos, pela agradável convivência. A Cynthia Paula Andrade pela amizade surgida no decorrer de interessantes colaborações científicas. A Sra. Ana Paula Oliveira, bibliotecária do ICB, pela atenção e carinho demonstrados nos processos de comutação, permitindo a aquisição de trabalhos científicos imprescindíveis para a revisão de literatura realizada no presente estudo. Aos colegas de turma de mestrado pelos saudosos momentos compartilhados durante a realização das disciplinas. Sem dúvida aprendemos muito uns com os outros. Ao Programa de Pós Graduação em Parasitologia, ICB, UFMG, em nome do atual coordenador Prof. Dr. Marcos Horácio Pereira, pela oportunidade de ser, com muito orgulho, discente deste departamento. A todos os professores do curso de Pós-graduação em Parasitologia, especialistas em suas respectivas áreas, que muito contribuíram para a minha formação parasitológica. A Sra. Sumara Aparecida Guilherme Ferreira, secretária da Pós-graduação em Parasitologia, pelo carinho maternal e serenidade na condução dos alunos do curso pelo melhor caminho possível. Ao Conselho Nacional de Pesquisa (CNPq) pela concessão da bolsa de Mestrado. Por fim, aos invertebrados estudados na presente dissertação, que através de complexas interações biológicas vem me demonstrando a simplicidade da vida. A todos vocês, os meus sinceros agradecimentos. 7 RESUMO Visando avaliar aspectos biológicos e taxonômicos de trematódeos transmitidos por Melanoides tuberculata (Müller, 1774) no Brasil, coletas malacológicas foram realizadas na represa da Pampulha, Belo Horizonte, Minas Gerais, no período entre março de 2006 a julho de 2009. De 3834 exemplares de M. tuberculata coletados e examinados, 279 (7,3%) apresentaram-se infectados por larvas de trematódeos que, após caracterização morfológica preliminar, foram utilizadas para estudos biológicos experimentais. Estágios evolutivos (cercárias, metacercárias, rédias, parasitos adultos, ovos e/ou miracídios) foram estudados a fresco e após processo de fixação, coloração e montagem de lâminas permanentes, seguida de análise morfológica e morfométrica em microscópio de luz. Cercárias do tipo Pleurolofocerca foram encontradas em 268 (7,0%) exemplares de M. tuberculata e utilizadas para a infecção de Poecilia reticulata Peters, 1859. Metacercárias obtidas nas brânquias dos peixes 30 dias após a infecção foram inoculadas por via oral a Mus musculus (Linnaeus, 1758) da linhagem AKR/J e parasitos adultos recuperados no intestino delgado dos camundongos 30 dias após infecção foram identificados como Centrocestus formosanus (Nishigori, 1924). Outro tipo cercariano, do grupo Megalura, foi encontrado em 11 (0,3%) exemplares de M. tuberculata coletados. Estas larvas formaram metacercárias em substrato sólido que foram utilizadas para a infecção de Gallus gallus domesticus (Linnaeus, 1758) jovens. Parasitos adultos obtidos da membrana nictitante e saco conjuntival das aves, 30 dias após infecção, foram identificados como Philophthalmus gralli Mathis & Leger, 1910. Este é o primeiro registro de ambas as espécies de trematódeos em M. tuberculata no Brasil. Palavras-chave: Melanoides tuberculata, trematódeos, Centrocestus formosanus, Philophthalmus gralli, infecção experimental, represa da Pampulha, Brasil 8 ABSTRACT With the aim to evaluate some biological and taxonomic aspects of trematodes transmitted by Melanoides tuberculata (Müller, 1774) in Brazil, malacological surveys were carried out at Pampulha dam, Belo Horizonte, Minas Gerais State from March 2006 to July 2009. From 3,834 specimens of M. tuberculata collected and examined, 279 (7.3%) were found infected by larvae of trematodes. After preliminary morphological characterization, larvae were used for experimental life cycle studies. Developmental stages (cercariae, metacercariae, rediae, adult parasites, eggs and/or miracidium) were studied alive and after fixation, staining and permanent mounting. The morphological and morphometric analyses were made using a light microscopy. Pleurolophocerca cercaria was found in 268 (7.0%) specimens of M. tuberculata and was used for experimental infection of laboratory reared Poecilia reticulata Peters, 1859. Metacercariae obtained from gills of fishes 30 days after the infection were inoculated by oral route in AKR/J strain of Mus musculus (Linnaeus, 1758). Adult parasites recovered in small intestine of mice, 30 days after the infection, were identified as Centrocestus formosanus (Nishigori, 1924). Another cercarian type, Megalura group, was found in 11 (0.3%) M. tuberculata specimens collected. These larvae encysted in solid substrate and were used for experimental infection of young chicken Gallus gallus domesticus (Linnaeus, 1758). Adult parasites recovered from the nictitant membrane and conjunctival sac of the chickens, 30 days after the infection, were identified as Philophthalmus gralli Mathis & Leger 1910. This is the first report of both fluke species in M. tuberculata from Brazil. Keywords: Melanoides tuberculata, trematodes, Centrocestus formosanus, Philophthalmus gralli, experimental infection, Pampulha dam, Brazil 9 LISTA DE ABREVIATURAS A: Aves L: Largura An: Anfíbios O: Ovos Bc: Bolsa do cirro Ov: Ovário C: Comprimento Pf: Pré-faringe Ce: Cecos intestinais Pg: Primórdio genital Ci: Cirro R: Répteis DPI: Dias pós-infecção M: Mamíferos E: Exensão N: Número Ea: Espinhos aciculares Ut: Útero EAU: Emirados Árabes Unidos T: Tipo Es: Esôfago Ta: Testículo anterior EUA: Estados Unidos da América Td: Testículo direito Fa: Faringe Te: Testículo esquerdo Gv: Glândulas vitelínicas Tp: Testículo posterior GPS: Global Positioning System Ve: Vesícula excretora ICG: Índice Cercariano Global Vo: Ventosa oral ICE: Índice Cercariano Específico Vs: Vesícula seminal Vv: Ventosa ventral LISTA DE FIGURAS FIGURA 1 - Melanoides tuberculata (Müller, 1774). Exemplar coletado na represa da Pampulha, Belo Horizonte, Minas Gerais................................................................15 FIGURA 2 - Croqui representativo da represa da Pampulha, Belo Horizonte, Minas
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